Novel Target For Immunotherapy Research Articles

Generation, Characterization, and Preclinical Studies of a Novel NKG2A-Targeted Antibody BRY805 for Cancer Immunotherapy.

Immuno-oncology has revolutionized cancer treatment, with NKG2A emerging as a novel target for immunotherapy. The blockade of NKG2A using the immune checkpoint inhibitor (ICI) monalizumab has been shown to enhance the responses of both NK cells and CD8+ T cells. However, monalizumab has demonstrated limited efficacy in in vitro cytotoxic assays and clinical trials. In our study, we discovered and characterized a novel anti-NKG2A antibody, BRY805, which exhibits high specificity for the human CD94/NKG2A heterodimer complex and does not bind to the activating NKG2C receptor. In vitro cytotoxicity assays demonstrated that BRY805 effectively activated NK92 cells and primary NK cells, thereby enhancing the cytotoxic activity of effector cells against cancer cells overexpressing HLA-E, with significantly greater efficacy compared to monalizumab. Furthermore, BRY805 exhibited synergistic antitumor activity when combined with PD-L1 monoclonal antibodies. In a mouse xenograft model, BRY805 showed superior tumor control relative to monalizumab and demonstrated a favorable safety profile in non-human primate studies.

Tumor Immune Microenvironment in Intrahepatic Cholangiocarcinoma: Regulatory Mechanisms, Functions, and Therapeutic Implications.

Treatment options for intrahepatic cholangiocarcinoma (iCCA), a highly malignant tumor with poor prognosis, are limited. Recent developments in immunotherapy and immune checkpoint inhibitors (ICIs) have offered new hope for treating iCCA. However, several issues remain, including the identification of reliable biomarkers of response to ICIs and immune-based combinations. Tumor immune microenvironment (TIME) of these hepatobiliary tumors has been evaluated and is under assessment in this setting in order to boost the efficacy of ICIs and to convert these immunologically "cold" tumors to "hot" tumors. Herein, the review TIME of ICCA and its critical function in immunotherapy. Moreover, this paper also discusses potential avenues for future research, including novel targets for immunotherapy and emerging treatment plans aimed to increase the effectiveness of immunotherapy and survival rates for iCCA patients.

Neudesin regulates dendritic cell function and antitumor CD8+ T cell immunity

Dendritic cells (DCs) are essential for antitumor T-cell responses to immune checkpoint inhibitor therapies. We have previously reported that the secreted protein neudesin suppresses DC function. In contrast, neudesin has been found to be abundantly expressed in human cancers. In this study, we evaluated the role of neudesin in cancer immunity. Cancer-related database analysis revealed that patients with melanoma with low neudesin expression exhibited increased infiltration of DCs and CD8+ T cells and improved outcomes of checkpoint inhibitor therapy. In mouse tumor models, neudesin deficiency delayed tumor growth and increased the proportions of Type 1 conventional DCs (cDC1s) and tumor antigen-specific CD8+ T cells in tumors and tumor-infiltrating lymph nodes. Neudesin-deficient antitumor cDC1 vaccine enhanced the systemic immunity more effectively than the wild-type cDC1 vaccine. Overall, our findings highlight the importance of neudesin in cancer immunity, providing a novel target for immunotherapy.

Subtype-specific alternative splicing events in breast cancer identified by large-scale data analysis

Genome analysis in cancer has focused mainly on elucidating the function and regulatory mechanisms of genes that exhibit differential expression or mutation in cancer samples compared to normal samples. Recently, transcriptome analysis revealed that abnormal splicing events in cancer samples could contribute to cancer pathogenesis. Moreover, splicing variants in cancer reportedly generate diverse cancer antigens. Although abnormal splicing events are expected to be potential targets in cancer immunotherapy, the exploration of such targets and their biological significance in cancer have not been fully understood. In this study, to explore subtype-specific alternative splicing events, we conducted a comprehensive analysis of splicing events for each breast cancer subtype using large-scale splicing data derived from The Cancer Genome Atlas and found subtype-specific alternative splicing patterns. Analyses indicated that genes that produce subtype-specific alternative splicing events are potential novel targets for immunotherapy against breast cancer. The subtype-specific alternative splicing events identified in this study, which were not identified by mutation or differential expression analysis, bring new significance to previously overlooked splicing events.

Abstract PO5-16-02: Investigating survivin as a novel target for immunotherapy in diverse breast cancer patients

Abstract Background: The BIRC5 gene encodes survivin, a tumor-specific antigen associated with aggressive tumor features, Black race and poor prognosis in breast cancer (BC). Recent studies have shown that anti-survivin vaccines stimulate a strong anti-tumor immune response in multiple tumor types. However, the relationship between BIRC5/survivin and immune response in BC is poorly understood, particularly among diverse patients. In this analysis, we sought to investigate the relationship between BIRC5/survivin and the BC immune microenvironment across clinical tumor features and RNA-based risk scores for genomic instability. Methods: We leveraged the Carolina Breast Cancer Study (CBCS), a large population-based study that oversampled young (≤50 years) and Black women with invasive BC. NanoString mRNA expression profiling was used to evaluate breast tumor expression for 10 major immune cell types, adaptive and innate immune response, TP53 and homologous recombination deficiency (HRD) pathways, and BIRC5/survivin from 1952 BC patients, including 1,030 (53%) Black and 922 (47%) non-Black women from the CBCS, with additional comparison in The Cancer genome Atlas (TCGA) (n=1,095 breast tumors). BIRC5 was evaluated both as a continuous and categorical variable, using the third expression quartile as a cut point for BIRC5-high, with the lower three quartiles categorized as BIRC5-low. We used generalized linear models to estimate beta values and adjusted p-values as the measure of association between BIRC5 status and immune-related expression in strata defined by estrogen receptor (ER) status, tumor stage, TP53 and HRD status. All analyses were adjusted for multiple comparisons with the Benjamini-Hochberg procedure. Results: In an analysis of continuous BIRC5 expression and global classes of immune response, BIRC5 mRNA expression was highest among tumors with an adaptive-enriched immune class (p< 0.001) in both CBCS and TCGA. However, when investigating immune gene expression according to BIRC5 status (i.e., BIRC5-high relative to BIRC5-low), multivariate differential expression analysis revealed significantly decreased immune cell-related expression scores among BIRC5-high tumors relative to BIRC5-low, and this relationship persisted in analysis stratified on stage (I/II vs III/IV), ER status, TP53 and HRD status. Across all strata, the strongest relationship between BIRC5-high and the immune microenvironment was observed for adaptive immune response (Beta: -0.14; p=0.005), with decreased immune gene expression related to B cells (Beta: -0.16; p=0.01), T cells (Beta: -0.14; p=0.007), Cytotoxic cells (Beta: -0.14; p=0.005), and PD-L1 (Beta: -0.15; p=0.005), relative to BIRC5-low. Conclusion: High BIRC5/survivin expression is associated with decreased immune-related gene expression in both ER-positive and ER-negative disease, and across all tumor stages. Anti-survivin therapies may be beneficial for boosting immune response in breast tumors with low immunogenicity, offering promise for BC subtypes previously ineligible for immunotherapy. Citation Format: Alina Hamilton, Qichen Wang, Sarah Soppe, Melissa Troester, Yara Abdou. Investigating survivin as a novel target for immunotherapy in diverse breast cancer patients [abstract]. In: Proceedings of the 2023 San Antonio Breast Cancer Symposium; 2023 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2024;84(9 Suppl):Abstract nr PO5-16-02.

Tumor suppressor function of RBMS3 overexpression in EOC associated with immune cell infiltration

ObjectivesEpithelial ovarian cancer (EOC) is considered to be a prevalent female malignancy with both high incidence and mortality. It is reported that RNA-binding protein 3 (RBMS3) executives a tumor suppressor function in different cancers. This investigation was designed to examine the expression of RBMS3 in epithelial ovarian cancer, the effects on EOC cells, and its connection to immune cells that infiltrate tumors in the EOC microenvironment. MethodsThe expression levels of RBMS3 in EOC tissues as well as their correlations with immune cell infiltration and clinical outcome were examined using bioinformatics approaches. Western blotting as well as immunohistochemistry were carried out to determine the protein levels in EOC tissues. In addition, qRT-PCR was employed to look at the expression of the mRNA. The role of RBMS3 in EOC cells was investigated, and an RBMS3 lentiviral vector was developed. The effects of RBMS3 on subcutaneous tumor development, the proliferation protein Ki-67, the tumor angiogenesis indicator CD31, and its function in controlling the tumor immune microenvironment were evaluated by in vivo tests. ResultsThere was a considerable decrease in RBMS3 expression in EOC tissues, which was linked to a poor prognosis for patients and the infiltration of multiple immune cell. Given immunohistochemical studies, tissues with increased RBMS3 expression had decreased markers of myeloid-derived suppressor cells, regulatory T cells, and M2 macrophages, whereas M1 macrophage markers were elevated. RBMS3 appears to suppress the capabilities of proliferating, invading, and migrating in EOC cells according to in vitro tests, whereas tumors overexpressing RBMS3 developed more slowly in syngeneic mouse models. The overexpression of RBMS3 led to a decline in the levels of Ki-67 protein and CD31. Additionally, it showed a negatively correlation with markers of regulatory T cell, myeloid-derived suppressor cell, and M2 macrophage but a positive correlation with markers of M1 macrophage. ConclusionsThe findings revealed that elevated RBMS3 expression plays a tumor suppressor role in EOC and was connected to patient survival in EOC. The studies conducted in vitro and in vivo demonstrated a link between RBMS3 expression and the infiltration of certain immune cells, indicating a function for RBMS3 in the immunosuppressive tumor microenvironment and its promising efficiency as a novel target for immunotherapy against EOC.

Heterogeneity of Lipid Metabolism and its Clinical and Immune Correlation in Lung Adenocarcinoma.

The role of lipid metabolism in lung adenocarcinoma (LUAD) is not completely researched. Lipid metabolism reprogramming is a characteristic of malignancies and contributes to carcinogenesis and progression. The transcriptome and scRNA- seq data and clinical information were downloaded from the public databases. Lipid metabolism pathways were collected from the MSigDB database, and molecular subtypes were classified based on lipid metabolism features via consensus clustering. The bidirectional crosstalk between immune cells and malignant cells was analyzed. Differences in lipid metabolism at the single-cell level and their correlation with the tumor microenvironment (TME) were also studied. LUAD patients were classified into two subtypes, showing distinct mutation and lipid metabolism features based on lipid metabolism characteristics. Meanwhile, significant differences in the overall survival, clinical characteristics, and immune landscape were observed between the two subtypes. We also found that clust1 had higher oxidative stress status. There were 116 differentially expressed genes between the two subtypes, which were significantly associated with cell cycle progression. We identified 4001 immune cells, including 483 malignant cells and 3518 normal cells, and found active intercellular communication and significant differences in lipid metabolism characteristics between the malignant cells and normal cells. Furthermore, several lipid metabolism pathways were found to be associated with TME factors, including hypoxia and angiogenesis. The current findings indicated that lipid metabolism was involved in the development and cellular heterogeneity of LUAD and revealed widespread reprogramming across multiple cellular elements in the TME of LUAD. This characterization improved the current understanding of tumor biology and enabled the identification of novel targets for immunotherapy.

Abstract 1373: Gene expression correlation of immune checkpoint molecules Siglec-15 and PD-L1 varies widely by cancer indication

Abstract Siglec-15 is a novel target for immunotherapy in cancer and exhibits low mRNA expression in normal tissues but broad expression across cancer indications, specifically on tumor-associated macrophages and tumor cells [1]. Expression of Siglec-15 and the immune checkpoint molecule PD-L1 have previously been reported to be mutually exclusive [1][2]. However, a large-scale meta-analysis of this claim has not been undertaken across cancer indications. Here, RNA-Seq analyses were performed across single cells, cell lines, and bulk tumor samples for Siglec-15 and PD-L1 mRNA expression. Data and meta-data from TCGA, CCLE, and primary tumor samples from 62 studies were obtained through OmicSoft OncoLand and single cell analysis was performed on the BioTuring Talk2Data platform. Analyses were limited to non-small cell lung cancer (NSCLC), cholangiocarcinoma, breast, thyroid, head and neck, colon, rectal, bladder, kidney, and endometrial cancers. Expression values were binned separately into percentiles across all indications and samples in increments of 10%. These analyses revealed individual gene expression levels of Siglec-15 and PD-L1 in bulk tumor samples varied substantially by indication, while co-expression of Siglec-15 and PD-L1 varied across indications. Analysis of gene expression of Siglec-15 and PD-L1 from the TCGA database showed a positive correlation in several indications including NSCLC, head and neck, kidney, and thyroid cancers. Analyses from additional studies in primary tumor samples confirmed co-expression of Siglec-15 and PD-L1 in NSCLC and head and neck cancer. Cell lines showed a similar trend of positive correlation with varying levels of Siglec-15 and PD-L1 expression. However, when examining expression at the single cell level (not bulk tumor), in 30 single cell studies spanning >1.3M cells, only 731 cells in 20 studies were found to co-express Siglec-15 and PD-L1 on the same cell while each gene was expressed individually on other cells. The majority of the cells co-expressing the genes were macrophages (n=~350), followed by CD4 (n=~115) and CD8 T cells (n=~50). This study demonstrates that at the bulk tumor level, Siglec-15 and PD-L1 mRNA expression are not broadly mutually exclusive across cancer indications and instead expression of each gene varies broadly within a given indication. Additionally, their expression is frequently positively correlated, including in NSCLC, head and neck, kidney, and thyroid cancers. These results also demonstrate that Siglec-15 and PD-L1 are rarely found co-expressed on same cell. These associations enable a better understanding of the landscape of target expression in patients across a wide variety of cancer indications and can inform combination strategies with anti-Siglec-15 therapies. [1] Sun J, et al. Clin Cancer Res, 2021;27(3):680-688. [2] Wang J, et al. Nat Med, 2019;25(4):656-666. Citation Format: Krystal Watkins, Liyang Diao, Marsha Crochiere, Jan Pinkas. Gene expression correlation of immune checkpoint molecules Siglec-15 and PD-L1 varies widely by cancer indication [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 1373.

Abstract A084: AMH paracrine signaling between cancer and mesothelial cells modulates the ovarian tumor immune microenvironment

Abstract A084: AMH paracrine signaling between cancer and mesothelial cells modulates the ovarian tumor immune microenvironment

Estrogen Receptor Mutations as Novel Targets for Immunotherapy in Metastatic Estrogen Receptor-positive Breast Cancer.

Estrogen receptor (ESR1) mutations have emerged as a key factor in endocrine therapy resistance. We identified and validated five novel, immunogenic ESR1-derived peptides that could be targeted through vaccine-based immunotherapy.

Targeting PD-1/PD-L1 inhibits rejection in a heterotopic tracheal allograft model of lung transplantation.

Immune checkpoint molecules such as programmed death-1 (PD-1) and programmed death ligand-1 (PD-L1) have revolutionized the field of lung cancer treatment. As part of our study, we examined the role of these proteins in acute rejection in a mouse model of heterotopic tracheal transplantation. Recipient mice were untreated (Allo group) or treated with anti-PD-L1 (aPDL1 group) or PD-L1 Fc recombinant protein (PD-L1 Fc group). A further group of C57BL/6 mice received isografts (Iso group). The occlusion rate was significantly higher in the Allo group than in the Iso group (p = 0.0075), and also higher in the aPD-L1 group (p = 0.0066) and lower in the PD-L1 Fc group (p = 0.030) than in the Allo group. PD-L1 Fc recombinant protein treatment significantly decreased interleukin-6 and interferon-γ levels and reduced the CD4+/CD8+ T cell ratio, without increasing PD-1 and T-cell immunoglobulin mucin 3 expression in CD4+ T cells. These data suggest that PD-L1 Fc recombinant protein decreases the levels of inflammatory cytokines and the proportion of CD4+ T cells without exhaustion. The PD-L1-mediated immune checkpoint mechanism was associated with rejection in the murine tracheal transplant model, suggesting a potential novel target for immunotherapy in lung transplantation.

CD155 on Multiple Myeloma Cells Is Epigenetically Regulated, Reduce Patients Survival and Compromises the Action of Novel Immunotherapies

Epigenetics, especially DNA methylation, is a key molecular regulator of gene expression. This regulation is important for cytotoxic cell activation (i.e., T-cells and NK cells) and is crucial for the clearance of malignant cells. Immune checkpoint (IC) events mediate cytotoxicity, one IC receptor involved in the process is CD155. Such receptor interacts primarily with TIGIT on cytotoxic cells and induces the inhibition of cytotoxicity. Multiple myeloma (MM) represents the 10% of all hematological malignancies and it has been described that MM cells hinder cytotoxic cell activation by inhibition and exhaustion. Furthermore, novel immunotherapeutic strategies such as Bispecific T-cell Engagers (BiTE) and Chimeric Antigen Receptor (CAR) therapies are being evaluated in MM treatment, with promising results. Nevertheless, they will confront a hostile environment. This is a very active field with frequent descriptions of new targets and therapeutic approaches being studied each day. We investigated whether CD155 is epigenetically regulated in MM and its importance in inducing inhibition of anti-tumor response in the context of novel immunotherapies through the CD155/TIGIT axis. To achieve our objectives, we focused on the promoter region of CD155 (the region up to 1500 base pairs upstream of the TSS) and characterized it at DNA, RNA and protein levels through bisulfite sequencing, quantitative PCR, and flow cytometry. We confirmed CD155 as an epigenetically regulated gene, thus demonstrating an existing correlation between CD155 promoter methylation and gene expression. In vitro treatment with azacytidine was performed to assess the active repression of CD155 expression by DNA methylation. Six different MM cell lines were characterized (two not expressing: AMO-1, KMS-12-BM, four expressing: RPMI-8226, MM.1S, EJM and JJN-3). From there, we created CD155 depletion models on expressing cell lines by short hairpin RNA interference and developed an in vitro coculture system with T-cells (from healthy donors). We observed a loss of expression between the 60% to 90% (when compared with controls). Depletion of CD155 did not significantly impact cell growth, apoptosis, or cell cycle. On the coculture with healthy T-cells, more cytotoxicity was detected against CD155-depleted cells (in RPMI-8226 Mann-Whitney P=0.020; JJN-3 and EJM P<0.001). To incorporate novel immunotherapeutic strategies, we carried out the same cocultures with 100pg/mL of anti-BCMA/CD3 BiTE, against anti-BCMA CAR-T or anti-BCMA CAR-NK cells (for RPMI-8226 and JJN-3 Mann-Whitney P=0.008). Coculture combinations were also performed in the presence of 10μg/ml of neutralizing αTIGIT (RPMI-8226, Mann-Whitney P =0.005; JJN-3 P=0.004) and/or αPD1 antibody. When novel immunotherapy approaches were included in the system, we observed how the depletion of CD155 was related to a cytotoxicity increase in all cases. In the presence of neutralizing αTIGIT, cytotoxicity was increased against CD155-expressing cells, while we observed no difference against CD155-depleted cells. In the presence of αPD1, we saw a significant restoration of T-cell cytotoxicity against both models and the antibody combination showed a synergic effect in CD155 expressing cells. Finally, we analyzed the existence of a correlation between CD155 gene expression and survival in newly diagnosed MM samples from CoMMpass project public data (N= 793). These data supported our results as the newly diagnosed MM patients with higher expression of CD155 had shorter overall survival than lower expressing ones (Logrank test P<0.001). In summary, in the context of MM, the expression of CD155 is regulated by the methylation status of its promoter region. Expression of CD155 effectively reduces cytotoxic cell response, which is mediated by the interaction of CD155 and TIGIT. The addition of αTIGIT and αPD1 validated that the CD155 cytotoxic cell inhibition is mediated in part by TIGIT interaction, independently of PD1/PD-L1 axis. This interaction seems to impact patients treated with current therapies, since their outcome correlates with CD155 expression. Based on our results, novel immunotherapies focused on enhancing immune cytotoxicity will benefit from CD155/TIGIT axis blockade. These results warrant further investigation on CD155 as a biomarker and target for novel immunotherapies for MM and other malignancies.

Single-Cell Profiling of Tumor-Associated Neutrophils in Advanced Non-Small Cell Lung Cancer.

Neutrophils act as a non-negligible regulator in the initiation and progression of malignancies, playing bifacial roles in the process. Thus, to understand the heterogeneity of tumor-associated neutrophils (TANs) comprehensively in advanced non-small cell lung cancer (NSCLC) at single-cell resolution is necessary and urgent. We applied single-cell RNA-sequencing (scRNA-seq) to portray the subtype-specific transcriptome landscape of TANs in advanced NSCLC using nine freshly obtained specimens. The scRNA-seq data were further processed for pseudo-time analysis to depict the developmental trajectory of TANs. Meanwhile, the interplay between TANs and other cell types within tumor microenvironment (TME) was revealed by intercellular interaction analysis. Seven distinct TAN subtypes were defined, of which, the N3 cluster was considered inflammatory phenotype expressing genes encoding multiple chemotactic cytokines, and correlated with inferior overall survival, indicating that N3 might be a pro-tumorigenic TAN subtype. N1 and N5 clusters were considered to be well differentiated and mature neutrophils based on CXCR2 expression and pseudo-time patterns, and both accounted for relatively high proportions in lung adenocarcinoma. In addition, genes related to neutrophil differentiation were discovered. We also found that TAN subtypes interacted most closely with macrophages through chemokine signaling pathways within TME. Our study refined TAN subtypes and mapped the transcriptome landscape of TANs at single-cell resolution in advanced NSCLC, collectively indicating the heterogeneity of TANs in NSCLC. Neutrophil differentiation- and maturation-related genes were also discovered, which shed light on different functions of TAN subclones in tumor immune escape, and may further provide novel targets for immunotherapy.

Ferroptosis and WDFY4 as novel targets for immunotherapy of lung adenocarcinoma.

Lung cancer exhibits the world's highest mortality rate among malignant cancers worldwide, thereby presenting a significant global challenge in terms of reducing patient mortality. In the field of oncology, targeted immunotherapy has emerged as a novel therapeutic approach for lung cancer. This study aims to explore potential targets for immunotherapy in lung adenocarcinoma (LUAD) through the analysis of Ferroptosis Index (FPI) and Single Cell RNA-Sequencing (scRNA-seq) data. The findings of this research can potentially offer valuable insights for improving LUAD immunotherapy strategies and informing clinical decision-making. Firstly, the relationship between survival and ferroptosis in LUAD patients was analyzed by FPI. Subsequently, the association between ferroptosis and infiltration and regulation of immune cells was explored by immune infiltration analysis and correlation statistics. Lastly, the relationship between major infiltrating immune cell populations and related pathways and prognosis of LUAD patients was analyzed by GSEA and GSVA. To screen out core genes regulating infiltration of immune cell populations, scRNA-seq data of cancer and para-cancerous tissues of LUAD patients were downloaded, followed by cell clustering analysis, cell identification of core subpopulations, pseudotime analysis, single-cell GSVA and pathway enrichment analysis, and identification and functional analysis of core regulatory genes. Moreover, the expression levels of core functional genes in LUAD tissue microarray were detected by immunohistochemistry, and its relationship with the prognosis of LUAD patients was verified. Finally, we used lentivirus with WDFY4 to transfect LUAD A549 cells. CCK-8, flow cytometry apoptosis detection, Scratch wound healing assay, Transwell migration assay, Xenograft nude mice model, immunohistochemical analysis and other experimental methods were used to explore the biological effects of WDFY4 on LUAD in vitro and in vivo. Survival analysis of FPI values in LUAD patients revealed a positive correlation between smaller FPI values and longer overall survival. Immuno-infiltration analysis and its correlation with FPI values revealed that B cells were most strongly associated with ferroptosis. Ferroptosis of cancer cells could promote infiltration and activation of B cell populations, and LUAD patients with more infiltration of B cell populations had longer long-term survival. scRNA-seq data analysis indicated that the B cell population is one of the major cell populations infiltrated by immune cells in LUAD. During the later phases of B cell differentiation in LUAD, there was a decrease in the expression levels of ACAP1, LINC00926, TLR10, MS4A1, WDFY4, and TRIM22 genes, whereas the expression levels of TMEM59, TP53INP1, and METTL7A genes were elevated. The protein-protein interaction (PPI) network analysis indicated that WDFY4 plays a crucial role in regulating B cell differentiation in LUAD. Immunohistochemical analysis of LUAD tissue microarray revealed a significant downregulation of WDFY4 expression, which was closely related to the occurrence sites of LUAD. Moreover, LUAD patients with a low WDFY4 expression exhibited a poorer prognosis. Additionally, experimental findings demonstrated that the overexpression of WDFY4 could inhibit the proliferation and metastasis of A549 cells while promoting apoptosis. It was also confirmed that WDFY4 could inhibit cancer growth in vivo. The results indicate that promoting infiltration and activation of B cell populations could improve the long-term survival of LUAD patients, thereby offering a potential novel immunotherapeutic approach for LUAD. Besides, the promotion of cancer cell ferroptosis and upregulation of WDFY4 expression have been shown to induce the infiltration and activation of B cell populations. Furthermore, the overexpression of WDFY4 can significantly inhibit the growth of lung adenocarcinoma in vitro and in vivo, highlighting its potential as a target for immunotherapy in LUAD.

Exploring the Role of DKK1 in the Occurrence of Lung Adenocarcinoma Based on the Analysis of Bioinformatics

Lung cancer is the most common malignant tumor in China, lung adenocarcinoma (LUAD) is the main type of lung cancer, which is a serious threat to people's life and health. At present, there are fewer studies on the role of Dikkopf1 (DKK1) in lung adenocarcinoma. The aim of this study was to investigate the role and potential prognostic value of DKK1 in the development of lung adenocarcinoma by bioinformatics methods. Several databases, such as genotype-tissue expression (GTEx), The Cancer Genome Atlas (TCGA) and tumor-immune system interactions database (TISIDB), were used to analyze the expression, clinicopathological features, immune cell infiltration, prognosis and methylation of DKK1 in lung adenocarcinoma. Then, linked immune cell infiltration Omics database was used to analyze the co-expressed genes of DKK1 and their functional enrichment. Finally, 59 pathological samples of paraffin-embedded lung adenocarcinoma patients who underwent surgery at the Affiliated Cancer Hospital of Xinjiang Medical University between 2016 and 2017 were collected for the validation of the prognostic value of expression by immunohistochemistry (IHC) test. The results of bioconfidence analysis showed that the expression level of DKK1 in lung adenocarcinoma tissues was higher than that in normal tissues, the expression in advanced cancers was higher than that in early stages, and the experimental validation revealed that among 59 cases of lung adenocarcinoma, there were 15 cases of negative expression (25.4%), 18 cases of weakly positive expression (30.5%), and 26 cases of strongly positive expression (44.1%). The different expression of DKK1 is related to methylation, prognosis and the activities of various immune cells. Functional enrichment shows that DKK1 may be involved in skin development and cell-substrate junction, and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis shows that DKK1 is related to ABC transporters. Bioinformatics analysis and clinical case specimens showed that high DKK1 expression was associated with poorer prognosis in patients with lung adenocarcinoma. High expression of DKK1 in lung adenocarcinoma is associated with poor prognosis. DKK1 is closely associated with tumor immune cell infiltration and pathways. DKK1 can be considered as a potential prognostic marker and a novel target for immunotherapy of lung adenocarcinoma.

Exploring the regulatory role of lncRNA in cancer immunity.

Imbalanced immune homeostasis in cancer microenvironment is a hallmark of cancer. Increasing evidence demonstrated that long non-coding RNAs (lncRNAs) have emerged as key regulatory molecules in directly blocking the cancer immunity cycle, apart from activating negative regulatory pathways for restraining tumor immunity. lncRNAs reshape the tumor microenvironment via the recruitment and activation of innate and adaptive lymphoid cells. In this review, we summarized the versatile mechanisms of lncRNAs implicated in cancer immunity cycle, including the inhibition of antitumor T cell activation, blockade of effector T cell recruitment, disruption of T cell homing, recruitment of immunosuppressive cells, and inducing an imbalance between antitumor effector cells (cytotoxic T lymphocytes, M1 macrophages, and T helper type 1 cells) versus immunosuppressive cells (M2 macrophages, T helper type 2 cells, myeloid derived suppressor cells, and regulatory T cells) that infiltrate in the tumor. As such, we would highlight the potential of lncRNAs as novel targets for immunotherapy.

HOPX is a tumor-suppressive biomarker that corresponds to T cell infiltration in skin cutaneous melanoma

BackgroundSkin cutaneous melanoma (SKCM) is the most threatening type of skin cancer. Approximately 55,000 people lose their lives every year due to SKCM, illustrating that it seriously threatens human life and health. Homeodomain-only protein homeobox (HOPX) is the smallest member of the homeodomain family and is widely expressed in a variety of tissues. HOPX is involved in regulating the homeostasis of hematopoietic stem cells and is closely related to the development of tumors such as breast cancer, nasopharyngeal carcinoma, and head and neck squamous cell carcinoma. However, its function in SKCM is unclear, and further studies are needed.MethodsWe used the R language to construct ROC (Receiver-Operating Characteristic) curves, KM (Kaplan‒Meier) curves and nomograms based on databases such as the TCGA and GEO to analyze the diagnostic and prognostic value of HOPX in SKCM patients. Enrichment analysis, immune scoring, GSVA (Gene Set Variation Analysis), and single-cell sequencing were used to verify the association between HOPX expression and immune infiltration. In vitro experiments were performed using A375 cells for phenotypic validation. Transcriptome sequencing was performed to further analyze HOPX gene-related genes and their signaling pathways.ResultsCompared to normal cells, SKCM cells had low HOPX expression (p < 0.001). Patients with high HOPX expression had a better prognosis (p < 0.01), and the marker had good diagnostic efficacy (AUC = 0.744). GO/KEGG (Gene Ontology/ Kyoto Encyclopedia of Genes and Genomes) analysis, GSVA and single-cell sequencing analysis showed that HOPX expression is associated with immune processes and high enrichment of T cells and could serve as an immune checkpoint in SKCM. Furthermore, cellular assays verified that HOPX inhibits the proliferation, migration and invasion of A375 cells and promotes apoptosis and S-phase arrest. Interestingly, tumor drug sensitivity analysis revealed that HOPX also plays an important role in reducing clinical drug resistance.ConclusionThese findings suggest that HOPX is a blocker of SKCM progression that inhibits the proliferation of SKCM cells and promotes apoptosis. Furthermore, it may be a new diagnostic and prognostic indicator and a novel target for immunotherapy in SKCM patients.

Identification of cuproptosis‐related subtypes and tumor microenvironment characteristics in colon cancer

AbstractCuproptosis is a form of regulated cell death, which is characterized by the lethal accumulation of excessive copper in cells. However, its role in colon adenocarcinoma (COAD) remains elusive. Our study aimed to decipher the biological function of cuproptosis‐related genes (CRGs) in patients with COAD. The expression data were obtained from The Cancer Genome Atlas, while the Gene Expression Omnibus database was used to verify the results. A total of eight differentially expressed CRGs were selected from patients with COAD. Subsequently, patients were stratified into three subtypes with distinct clinical and biological features. In view of the huge differences in the prognosis between subtypes A and C, we selected them for further study, including the variations in clinical progressions, oncogenic pathways, and immune cell infiltration. For the sake of better evaluation, we also established a cuproptosis index (CI) to quantify the heterogeneity of CRGs expression. Enrichment analysis showed that the high‐CI group was enriched in immune activation pathways. Meanwhile, the immunosuppressive cell infiltration and immune checkpoints were elevated in the high‐CI group. The CI we constructed had its predicting function in different clinical groups. Besides, we noticed that CRGs, especially CDKN2A, were closely related to the clinical progression in patients with COAD and immune cell infiltration in tumors. Moreover, CDKN2A could become a potential novel target for immunotherapy in the setting of COAD.

Bispecific anti-CD3×anti-CD155 antibody mediates T-cell immunotherapy in human haematologic malignancies.

T cells are important components in the cell-mediated antitumour response. In recent years, bispecific antibodies (Bi-Abs) have become promising treatments because of their ability to recruit T cells that kill tumours. Here, we demonstrate that CD155 is expressed in a wide range of human haematologic tumours and report on the ability of the bispecific antibody anti-CD3 x anti-CD155 (CD155Bi-Ab) to activate T cells targeting malignant haematologic cells. The specific cytolytic effect of T cells armed with CD155Bi-Ab was evaluated by quantitative luciferase assay, and the results showed that the cytolytic effect of these cells was accompanied by an increase in the level of the cell-killing mediator perforin. Moreover, compared with their unarmed T-cell counterparts, CD155Bi-Ab-armed T cells induced significant cytotoxicity in CD155-positive haematologic tumour cells, as indicated by lactate dehydrogenase assays, and these results were accompanied by increased granzyme B secretion. Furthermore, CD155Bi-Ab-armed T cells produced more T-cell-derived cytokines, including TNF-α, IFN-γ, and IL-2. In conclusion, CD155Bi-Ab enhances the ability of T cells to kill haematologic tumour cells, and therefore, CD155 may serve as a novel target for immunotherapy against haematologic malignancies.

ECM2, a prognostic biomarker for lower grade glioma, serves as a potential novel target for immunotherapy.

Extracellular matrix protein 2 (ECM2), which regulates cell proliferation and differentiation, has recently been reported as a prognostic indicator for multiple cancers, but its value in lower grade glioma (LGG) remains unknown. In this study, LGG transcriptomic data of 503 cases in The Cancer Genome Atlas (TCGA) database and 403 cases in The Chinese Glioma Genome Atlas (CGGA) database were collected to analyze ECM2 expression patterns and the relationship with clinical characteristics, prognosis, enriched signaling pathways, and immune-related markers. In addition, a total of 12 laboratory samples were used for experimental validation. Wilcoxon or Kruskal-Wallis tests demonstrated highly expressed ECM2 in LGG was positively associated with malignant histological features and molecular features such as recurrent LGG and isocitrate dehydrogenase (IDH) wild-type. Also, Kaplan-Meier (KM) curves proved high ECM2 expression could predict shorter overall survival in LGG patients, as multivariate analysis and meta-analysis claimed ECM2 was a deleterious factor for LGG prognosis. In addition, the enrichment of immune-related pathways for ECM2, for instance JAK-STAT pathway, was obtained by Gene Set Enrichment Analysis (GSEA) analysis. Furthermore, positive relationships between ECM2 expression with immune cells infiltration and cancer-associated fibroblasts (CAFs), iconic markers (CD163), and immune checkpoints (CD274, encoding PD-L1) were proved by Pearson correlation analysis. Finally, laboratory experiments of RT-qPCR and immunohistochemistry showed high expression of ECM2, as well as CD163 and PD-L1 in LGG samples. This study identifies ECM2, for the first time, as a subtype marker and prognostic indicator for LGG. ECM2 could also provide a reliable guarantee for further personalized therapy, synergizing with tumor immunity, to break through the current limitations and thus reinvigorating immunotherapy for LGG. AVAILABILITY OF DATA AND MATERIALS: Raw data from all public databases involved in this study are stored in the online repository (chengMD2022/ECM2 (github.com)).